ID 75668
Title Alternative
改良型pHプロ-ブによる細胞内pHの定量的な計測方法の開発
Author
Nakata, Eiji Division of Bioinformatics Engineering, Department of Life System, Institute of Technology and Science, Graduate School of the University of Tokushima
Yukimachi, Yoshihiro Graduate School of Advanced Technology and Science, The University of Tokushima
Nazumi, Yoshijiro Graduate School of Advanced Technology and Science, The University of Tokushima
Uto, Yoshihiro Division of Bioinformatics Engineering, Department of Life System, Institute of Technology and Science, Graduate School of the University of Tokushima Tokushima University Educator and Researcher Directory KAKEN Search Researchers
Hori, Hitoshi Division of Bioinformatics Engineering, Department of Life System, Institute of Technology and Science, Graduate School of the University of Tokushima
Keywords
Fluorescent pH indicator
bio-imaging
intracellular pH measurement
SNARF
Content Type
Departmental Bulletin Paper
Description
SNARF is one of the most commonly used pH indicators for biological applications, owing to its unique fluorescent characteristics. For intracellular applications, esterase-substrate derivatives of SNARF, such as acetate or acetoxymethyl esters, have been employed previously as a generally accepted strategy to increase cell permeability. Unfortunately such cell-permeable SNARF derivatives retain significant fluorescence in aqueous solution, a property which results in a low signal-to-noise ratio. This in turn can lead to incorrect intracellullar pH measurements. Here we describe UTX-40, a newly designed SNARF derivative that successfully addresses these problems. In aqueous solution, UTX-40 is devoid of fluorescence before ester hydrolysis because it exists in aqueous environment as nano-scaled aggregates. As UTX-40 is converted into SNARF by enzymatic hydrolysis inside the cell, the aggregates become diffused and monomeric SNARF displays its characteristic fluorescent properties. The results of our studies reported in this communication demonstrate clearly the benefits of UTX-40 as an intracellular pH indicator. Since intracellular localized fluorescence was observed without cell-washing, the efficient uptake of intracellular fluorescence was confirmed. In addition, the actual intracellular pH and changes in intracellular pH caused by drug addition were monitored. The low background noise produced by UTX-40 is a property of this new pH probe that should be particularly advantageous for in vivo usage because, for this application, it is difficult to wash out the redundant probe.
Journal Title
徳島大学大学院ソシオテクノサイエンス研究部研究報告
ISSN
21859094
NCID
AA12214889
Volume
56
Start Page
45
End Page
50
Sort Key
45
Published Date
2011
FullText File
language
eng
departments
Bioscience and Bioindustry